用于蛋白质间相互作用研究的分析技术

概述近年来用于蛋白质间相互作用研究的分析技术,如酵母双杂交系统、荧光共振能量转移技术、表面等离子体共振技术、蛋白质芯片技术和生物信息学方法等,分别介绍其应用原理和特点,为蛋白质间相互作用研究及构建蛋白质相互作用图谱提供理论依据。     

表面等离子共振(SPR)技术在生物药物质量控制中的应用前景

摘要：表面等离子共振（SPR）技术是近年来飞速发展的一种新型光学分析技术,该技术在化学信号检测、生物药物分析、环境监测等多领域具有极大的应用价值。本文介绍了表面等离子共振技术的基本光学原理及常用的生物传感器,对表面等离子共振技术在生物药物分析和质量控制中的研究进展做一综述,并对该技术在生物药物质量控制领域的应用前景做一展望。     

表面等离子体共振生物传感器研究进展

本文介绍了一种新型的生物传感器——表面等离子体激元共振(SPR)传感器,它是进行生物分子相互作用分析的一种先进手段。与其他技术相比,它具有实时检测、无需标记、耗样量较少等优点,在药物筛选、Il缶床诊断、细胞膜模拟和生物大分子相互反应等领域中的新兴应用日益扩大。本文介绍了 SPR 的结构、原理、优点、应用及其新进展。随着联用技术和纳米技术的深入研究,SPR 将有广泛的应用前景。     

表面等离子共振技术研究进展及应用

表面等离子体共振(SPR)技术是自20世纪70年代逐步发展起来的一种光学生化检测技术。经过几十年的发展,因该技术具有样品免标记、需求量小、检测快速、灵敏度高、特异性强、选择性高等优势,已逐步发展成为分子生物学、环境污染物分析、食品安全检测、药物研发等领域常用的分析工具。本文系统综述了SPR技术发展的历史及在各个领域的应用情况,并展望了SPR技术未来发展的趋势。     

表面等离子共振技术在新药开发研究中的应用进展

表面等离子共振技术(SPR技术)在基因工程药物研究中的应用发展迅速。文章综述了近年来SPR技术在β-淀粉样蛋白(Aβ)沉积的保护蛋白质研究及抗癌新药和艾滋病病毒新抑制剂筛选中的应用进展。     

共振散射技术在痕量药物分析中的研究进展

本文评述了共振散射技术在痕量药物分析中的研究进展。分别综述了药物与染料、金属离子、无机纳米粒子、表面活性剂、β-环糊精、无机酸根、有机溶剂等相互作用以及药物的自凝聚作用,使共振散射显著增强,从而提高药物检测灵敏度。     

共振散射技术在痕量药物分析中的研究进展

本文评述了共振散射技术在痕量药物分析中的研究进展。分别综述了药物与染料、金属离子、无机纳米粒子、表面活性剂、β-环糊精、无机酸根、有机溶剂等相互作用以及药物的自凝聚作用,使共振散射显著增强,从而提高药物检测灵敏度。     

表面等离子共振技术在定量分析中的应用和研究

表面等离子共振（surface plasmon resonance,SPR）是一种基于物质间相互作用所导致的芯片表面质量变化而产生的一种光学现象,能够简单、快速、准确地对分子间作用的强度进行动态监测。SPR生物传感器具有无标记、灵敏度高、检测结果准确、可在线实时检测等优点。SPR技术在药物发现、临床诊断、食品安全、环境监测、蛋白组学等领域都有应用。对SPR在食品卫生、环境监测、临床应用中定量分析的相关应用和研究进行综述,期望能为相关研究提供参考。     

中药注射剂(类)致敏成分检测技术进展

中药注射剂易引起过敏反应,这是中药安全性评价研究的热点,也是亟待解决的难点之一。本文介绍了中药注射剂的不良反应概况,中药注射剂引起过敏反应的原因、类型;归纳了中药注射剂中的(类)致敏成分的主要检测技术,包括体外肥大细胞脱颗粒检测、细胞膜色谱技术、酶联免疫吸附实验、表面等离体子共振技术等,并分别介绍其原理、实验方法,局限不足处,并对比其适用范围及其在(类)致敏成分检测中的应用,以期为开展相关研究提供思路及参考。     

荧光共振能量转移技术及其在药物定量分析中的应用

荧光共振能量转移技术作为一种高效的光学“分子尺”,在生物医学、免疫分析以及核酸检测等方面已有广泛的应用,近年来又被用于药物的定量分析。综述荧光共振能量转移技术的原理和特点以及在药物定量分析中的应用。     

基于药物靶标识别中药活性成分的研究方法及应用

中药小分子与生物体内靶标蛋白的相互作用是中药发挥药理作用的基础，现代科学技术的发展和药物作用靶标的揭示为中药活性成分的研究提供了新的技术和手段。基于已知的药物作用靶标，阐明中药的有效成分及其作用机制已成为中药学研究和发展的重要方向。基于化学和生物技术与计算机虚拟筛选技术两大视角，总结了亲和超滤质谱技术、分子生物色谱技术、磁珠富集技术、等离子共振技术、生物膜干涉技术、分子对接技术、药效团模型和机器学习8种基于药物靶标识别中药活性成分的研究方法与应用现状，以期补充传统药物发现的方法，为该领域研究提供借鉴与参考。     

Research progress of surface plasmon resonance technology in drug discovery

Surface plasmon resonance (SPR) technology is a powerful and sensitive tool for investigating molecular interactions. Technical improvements in recent years have brought new functionality to SPR apparatuses that can be widely applied at multiplestages of the drug discovery process. The technology allows users to conduct ligand fishing and offers rich and in-depth information regarding the affinity, specificity, kinetics, concentration and identification of covalent/allosteric/competitive binding behaviors. The present review highlights the principle, sample types, detection ranges, experimental methods, strengths, limitations, and the latest research progress on the application of SPR technology in the drug discovery process.     

Biotin sulfone as a new tool for synthetic oligosaccharide immobilization: application to multiple analysis profiling and surface plasmonic analysis of anti-Candida albicans antibody reactivity against alpha and beta (1-->2) oligomannosides

As a part of our glycoantigen synthetic program for diagnosis and basic analysis of yeast-related pathogenic mechanisms, a library of 1-->2 oligomannosides suitable for immunoanalysis was prepared. The use of biotin sulfone, an oxidized form of biotin, offers a convenient solution for both oligosaccharide synthesis and immobilization on microspheres and surface plasmon resonance sensors. The application of this new strategy for the analysis of anti- Candida albicans antibody response through multiple-analyte profiling technology (Luminex) and with surface plasmonic analysis using biotin tagged synthetic oligosaccharides on avidin coated surfaces was validated using monoclonal antibodies.     

Biomolecular interaction analysis in drug discovery using surface plasmon resonance technology

The review gives first an introduction into the basics of surface plasmon resonance technology. The physical principle is shortly discussed followed by a discussion of the experimental details to be considered when using this technology for biomolecular interaction analysis. Based on recent publications it is demonstrated that the technology has widespread applications in many fields of the drug discovery process. Protein/protein interactions can be monitored in real time when working with biopharmaceuticals as well as protein/small analyte interactions during hit finding, secondary screening, lead optimization and lead selection. Equilibrium binding constants, kinetic rate constants and thermodynamic parameters are obtained from such study that helps to understand the mechanism of the binding reactions. This information can be directly used to improve binding properties of a drug candidate.     

Surface plasmon resonance in monitoring of complement activation on biomaterials

When artificial materials come into contact with blood, various biological responses are induced. For successful development of biomaterials used in biomedical devices that will be exposed to blood, understanding and control of these interactions are essential. Surface plasmon resonance (SPR) spectroscopy is one of the surface-sensitive optical methods to monitor biological interactions. SPR enables real-time and in situ analysis of interfacial events associated with biomaterials research. In this review, we describe an SPR biosensor and its application to monitor complement activation onto biomaterials surface. We also discuss the effect of surface properties of the material on complement activation.     

Bioanalytical Screening of Riboflavin Antagonists for Targeted Drug Delivery - A Thermodynamic and Kinetic Study

The present study screened riboflavin mimicking small molecules to determine their binding activity for the riboflavin binding protein. We performed thermodynamic and kinetic binding studies of these molecules using a combination of two analytical approaches; isothermal titration calorimetry and surface plasmon resonance spectroscopy. Screening of a biased set of non-riboflavin based small molecules by microcalorimetry led to the discovery of two known drug molecules, quinacrine and chloroquine, as favorable ligands for the riboflavin receptor with K(D) value of 264, and 2100 nM, respectively. We further demonstrated that quinacrine is a competitive ligand for the receptor as measured by surface plasmon resonance. Thus this study describes the identification of a novel class of dual acting riboflavin antagonists that target riboflavin receptor for cellular uptake and display multifunctional activities upon cellular entry.     

Implementing surface plasmon resonance biosensors in drug discovery

Recent improvements in instrument hardware, experimental design and data processing have made it possible to use surface plasmon resonance (SPR) biosensor technology in the discovery and development of small-molecule drugs. The key features of SPR biosensors (i.e. real-time binding analysis and lack of labeling requirements) make this technology suitable for a wide range of applications. Current instruments have a throughput of approximately 100-400 assays per day, providing a complement to secondary screening. The ability to collect kinetic data on compounds binding to therapeutic targets yields new information for lead optimization. Small-molecule analysis and emerging applications in the areas of ADME (adsorption, distribution, metabolism and excretion) and proteomics have SPR biosensors poised to play a significant role in the pharmaceutical industry.     

Single-step, label-free quantification of antibody in human serum for clinical applications based on localized surface plasmon resonance

The amount of antibody in blood is an important measure of health status for making critical decisions in clinical practice. Here, we demonstrated a single-step, label-free, molecular diagnostic method based on localized surface plasmon resonance (LSPR) using standard 96-well microtiter plates. We improved the LSPR biosensor so that it can measure antibodies to blood group antigens in human serum with a single-step operation. First, we employed the ampholytic polymeric surface modification technique to present an efficient molecular scaffold on the sensor surface. Second, we selected the combination of an appropriate reference molecule against the antigen and a blocking agent to significantly reduce the variability of signal due to nonspecific responses of the unknown in the sample. Finally, we overcame the analytical difficulty arising from serum and achieved a single-step "wash-free" measurement of the amount of target antibody in human serum. FROM THE CLINICAL EDITOR: In this paper, a novel, single-step, label-free, molecular diagnostic method is discussed for antibody detection based on localized surface plasmon resonance using standard 96-well microtiter plates.     

表面等离子共振技术测定药物与人血清白蛋白的相互作用

目的应用表面等离子共振技术(SPR)建立一种新的测定白蛋白与化合物相互作用的方法。方法将人血清白蛋白(HSA)固定在CM5芯片表面。不同化合物的溶液流经固定于芯片表面的HSA,SPR显示在芯片表面的折射系数变化。分析软件处理数据,用HSA流通池得到的数据减去参比池得到的数据进行拟合,确定结合常数。结果所有测试药物与固定的HSA可逆结合。紫草素1的平衡结合常数KA为3000L.mol-1。SPR样品需要量最小而且能够自动分析,还可以直接检测小分子(Mr308~585)结合,使之适用于评价药物与HSA相互作用。结论HSA主要的药物键合位点没有因为被固定到芯片表面而破坏。验证了SPR可以准确简易测定化合物的结合解离。